PB84-10013D
    Effects  of Selected Inorganic
    Coal-Gasification Constituents on
    Aquatic  Life:  An Annotated Bibliography
    Tennessee Valley Authority, Muscle Shoals,  AL
    Office  of Natural Resources
    Mar  83
U.S. Department of Commerce
National Technical Information Service

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                                  TECHNICAL REPORT DATA
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                                                 EPA-600/7-83-018
                                                 March 1983
    EFFECTS  OF SELECTED  INORGANIC COAL-GASIFICATION
             CONSTITUENTS ON AQ'JATIC  LIFE.:
               AN  ANNOTATED BIBLIOGRAPHY
                     Prepared by

                     Leroy M. Koch

                Principal Investigator
                   Richard C. Young

              Office  of  Natural Resources
              Division of Water Resources '.
          Fisheries & Aquatic Ecology Branch
              Tennessee  Valley Authority
             Muscle Shoals,  Alabama  ^5660
     Interagency Agreement  No.  EPA-IAG-79-0-X0511
                  Project No.  82  BDW
              Program Element  No.  CC2N1A
                    Project Officer

                     Alan Moghissi
          Office of Research and Development
         U.S.  Environmental Protection Agency
                 Washington, DC  20460
                     Prepared for

OFFICE OF ENVIRONMENTAL PROCESSES AND EFFECTS RESEARCH
          OFFICE OF RESEARCH AND DEVELOPMENT
         U.S.  ENVIRONMENTAL PROTECTION AGENCY
                 WASHINGTON, DC  2.0460

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                                 DISCLAIMER
          This report was prepared by the Tennessee Valley Authority and
has been reviewed by the Office of Research and Development, Energy and
Air Division, U.S. Environmental Protection Agency, and approved for
publication.  Although the research described in this document has been
funded wholly or in  part by the United States Environmental-'Protection
Agency through Interagency Agreement I!o. EPA-IAG-79-D-X0511 with TVA, it
has not been subject to Agency policy and peer review and therefore does
not necessarily reflect the views of the agency or the Tennessee Valley
Authority and no official endorsement should be inferred.
                                     ii

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                                  ABSTRACT

          This review is concentrated on primary inorganic pollutants of
concern which result in the aqueous discharges of"high-pressure coal-
gasification technologies.   These pollutants include ammonia, cyanide
(thiocyanates), sulfide, and boron.  Ammonia toxicity. was not reviewed
since effective waste treatment-technologies and discharge guidelines are
available.

          The open literature concerning the effects of cyanide (thiocyanates),
boron, ?nd sulfide on aquatic life was reviewed using computer search
technique?, to obtain information from the Department of Energy RECON data
base,, the BIOSIS biological file in ORBIT data base, chemical abstracts,
and the National Technical  Information Service.  Key words used-included
coal gasification, toxicity, teratogenesis,  aquatic organisms, fishes,
algae, Daphnia pulex, cyanide, thiocyanates, sulfide, sulfite, and boron.

          It was concluded  that there is i void in the literature concerning
the toxicity o  these parameters in high-pressure,  coal-gasification
wastewater matrices.  The information-abstracted .is mai.nly. concerned-with
laboratory studies with individual constituents; however, some data
provide insight to the effects-of—physical parameters such as temperature
and physical stress.
                                   iii

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                             ACKNOWLEDGEMENTS
          This  work  was  conducted  as-part  of  the Federal  Interagency
Energy/ Environment  Research  and Development  Program  with funds  administered
through the Environmental  Protection  Agency  (EPA Contract No.  79-D-X0511).

          The EPA Project  Officer  for the  project  is  Alan Moghlssi,  and
the TVA Project Director is  Billy  G.  Isom, E&D  Building,  Muscle  Shoals, Alabama.
                                   iv

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                                  CONTENTS
Abstract	      iii
Acknowledgement	       iv
                                   .                       )
CYANIDE	'	        1
     Fish  	......        2
        Eggs and Hatching	        2
        Sac c ry	        2
        Juvenile	        3
        Adults	.'  .  .  .        3
        Physiology	  .  .	        7
        Behavior	        9
        Bioconcentration	•	'.    .  .        9
     Invertebrates	       10
        Acute Effects	:  .  .  .       10
BORON	:	       11
     Fish    	•-.•-•       !2
        Acute and Chromic	       12
     Invertebrates	  .  .  .       15
        Acute	'.	    -...-,i5_
     Aquatic Plants	•.  .  .	       16
     Microorganisms   	       16
SULFIDE	i  .  .  .       17
     Fish	       18
        Eggs and Hatching	       18
        Sac Fry	•  •  •  •       19
        Juvenile -  Adults	       20
           Chronic  Tests  -  Fish-Bluegills	  .  .  .       21
        Physiology	-  -  - '	'.  .  .  .       26
     Invertebrates  	  ...;...       30
        Acute and Chronic	•.  .  .  .       30

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CYANIDE

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                                    FISH

                             (ACUTE AND CHROMIC)
EGGS AND HATCHING
Lind, David T.,  Lloyd L.  Smith, Jr., and Steven J. Broderius.  1977.
     Chronic effects of hydrogen cyanide on the fathead minnow.  Journal
     WPCF, 49:262-268.

          I.ind et al. (1977) found the number of eggs produced by female
fathead minnows was significantly reduced at concentrations of 0.0196
mg/1 HCN and above.


Leduc, Gerard.   1978.  Deleterious effects of cyanide on early life
     stages of Atlantic salmo.i (Salmo salar).   Journ. Fish. Res. bd.
     Canoda, 35:166-174.                                  :

          Leduc (1978)  found continuous exposure of Atlantic salmon eggs
to cyanide in a range of 0.01-0.10 mg/1 HCN caused hatching failures of
about 15 to 40%.

          Leduc (1978).  Hatching was delayed 6-9 days-at cyanide concentra-
tions of 0.08 and 0.10 mfi/1 HCN; water temperature was 5.4°C.  Hatching
was not delayed at lower concentrations of HCN.
Liiid, David T.,  Lloyd L. Smith, Jr., and Steven J. Broderius.  '1977.
     Chronic effects of hydrogen cyanide on tlie fathead minnow.  Journal
     WPCF, 4°:26^-268.

          Lind et al. (1977) estimated that tlie highest no-effect level
of hydrogen cyanide for the fathead minnow lies between 0.0129 and O.ol96
•fig/1 based on statistical evaluation of egg production.
SAC FRY - JUST AFTER HATCHING

Leduc, Gerard.  1978.  Deleterious effects of cyanide on early life
     stages of ..tlantic salm .1 (Salnic salar).   Journ. Fish. Res. Bd.
     Canada, 35:166-174.

          Ledur (1978) determined cyanide had no apparent effect on
survival of Atlantic salmon sac  fry after 'latching, but markedly affected
growth-producing fry that were 10% bhorte.r than controls at concentrationr.
of 0.04 mg/1 HCN and greater.

          Leduc (1978) noticed in Atlantic salmon fry an accelerated
increase in body length after hatching  in low concentrations of cyanide.

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          !n 0.08 and 0.10 mg/1  IICN,  58 -lays alter hati'hing, the fry,
initially smaller than the controls,  were equal  to if not longer than the
controls.  In addition, those fry exposed to intermediate concentrations
were longer than the controls.   Increase in weight essentially paralleled
the observations of increase in  length.

          Leduc (1978) found, during  the pcsthatching period, abnormal
Atlantic salmon fry were present with observable defects including mal-
formation and/or absence of the  eyes, defects in mouth and vertebral
column, and yolk-sac dropsy (hydrocoele embryonalis).

          Leduc (1978) showed a  high  incidence of abnormalities in Atlantic
salmon ranging from about 6% at  0.0-1  mg/I to about 19% at 0.10 mg/1 HCN
at a water temperature of 4.4°C."


Lind, David T., Lloyd L.  Smith,  Jr.,  and Steven J. Broderius.  1977.
     Chronic effects of hydrogen cyanide on the fathead minnow.  Journal
     WPCF, 49:262-268.

          Lind et al. (1977) found that fry exposed to treatment levels
of 34.8 pg/1 HCN and above after 28 days were significantly  shorter than
control  fish.  In one treatment  level belov 34.8.iig/l HCN at 26.3 pg/1
HCN, fish were significantly longer than controls.

          Lind et al. (1977) found that fathead minnow fry  in treatment
experiments with HCN after 28 and 56 days tended to approach the same
size of control fish.
JUVENILE

Lind, David T., Lloyd L. Smith, Jr., and Steven J. Broderius.   1977.
     Chronic effects of hydrogen cyanide on the fathead minnow.  Journal
     WPCF, 49:262-268.

          Lind et al. (1977) determined that che  lethal threshold concentra-
tion of hydrogen cyanide for juvenile fathead minnows at 25°C,  pH 8,  and
6 mg/1 DO, derived from acute toxicity bioassays, is approximately 0.012
mg/1.
ADULTS

Shelford, Victor E.   1917.  An experimental study of the effects  of  gas
     waste  upon fishes, with especial reference  to stream pollution.
     Bull.  Illinois State Laboratory Nat. History, Vol. XI, Article  VI,
     pp. 395-398.

          Shelford  (1917) found when Lepom_ij> huini l_u? were exposed to a
concentration of 0.280-0.300 grants of' ammonium sulphocyanate  (NH.j)

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 (280-300 ppm;  ;uid  a  concentration of ammonium  fcrrocyanide  (^(4)4,  O.liO-
 0.200 OID (.150-200  ppc)  spasms wore  common before: death which  occurred in
 one  hour .it  a  tcmperatjie of 20°C.


 Herbert, D.W.M.  and  J.  C. Merkens.   1952.  The  tcxicity  of  potassium
      cyanid; to  trout.  Jour. Exp.  Bio., 29:632-649.

           Herbert  aad Merkens (1952) showed  in  experiments  with  one-year-old
 rainbow trout, ranging  in length from 5.5 to 17.5  cm  in  dynamic  bioass^ys,
 water temperature  17.5, with a  concentration o£..0-l53--ppni--C:<,  that  longer
 fish usually succumbed  sooner than  shorter fish.


 Doudoroff, Peter,  Gerard Ledu'c, and Carl K. Schneider.   1966.  Acute
      toxicity  to fish of solutions  containing  complex metal cyanides, in
      .relation  to concentrations of  molecular hydrocyanic acid.   Trans.
      Amer. Fish. Soc.,  95:6-22.

           Doudoroff  et  al.  (1966) determined in tests of continually
 renewed NaCN solutions  at 20°C, with a pH of 7.8 to 7.9, thu free  cyanide
 concentrations near  and below 0.15  mg/1 as HCN  could  be  tolerated by raost
 of  the bluegills tested indefinitely.

           Doudoroff  et  al.  (1966).  The median  tolerance limits  (TL ) for
 bluegill is  about  0.14  or 0.15  mg/1 of molecular HCN  with a pH of 7T85.
 Lower concentrations were indefinitely tolerable for  bluegill unr'er test
 conditions.
. Broderius ,. Steven J.  and Lloyd  L.  Smith, Jr.   1979.   Lethal and sublethal
      effects of binary mixtures  of cyanide  and  hexa-valent chromium, zinc,
      or ammo'ni.a to the fathead  minnow  (Pimephales  promclas) and rainbow
      trout (Sal IRQ gairdneri) .   Journ.  Fish. Res.  Bd.  Canada, 36:164-172.

           Broderius and Smith  (19-9) determined the  96-hour r,Cso of HCN
 to fathead minnows and rainbow  trout to be  0.125  and  0.057 rag/1, respectively.
 Water temperature for the minnows  was  25°C  and  10°C  for the trout, while
 pH was 7.8 for the minnow tests  and 7.95 for  the  troLt.

           Broderius and Smith  (1979) determined 96-hour LCs0s,. in.wat /
 with temperature and pH as above,  for  fathead minnows in Cr-HCN to be
 1.31 toxic units and to rainbow trout  of Niis-IICN  to  be 0.859.. toxic units..
           Broderius and Smith  (1979).   In  30-day tests on fathead minnows
 with Zn at 0.76 and 0.34 nig/1  and  those with  a  mixture of HCN and Zn at
 0.045 and 0.62 and 0.057 and 0.86  mg/1, respectively, survival was about
 65% and 40% of that in the controls,  respectively.

           Erode'rius and Smith  (19/9).   Rainbow  trout were markedly affected
 by 0.044 mg/1 HCN with survival  after 30-day  exposure being 30% of that
 in the controls.

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Bu-dick, George Edgar and Morris LipschueLy.  5948.'  Toxicity of fcrro-
     and ferricyanide solutions to fish ;md determination of the cause of
     mortality.  Trans. Amer. Fish. So.:., 78:192-202.

          Burdick and Lipschuety (1948) found that a cyanide concentration
of approximately 0.19 ppm was toxic in laboratory tests to Rhinichthys
atratulus and Semotilus atromaculatus atromaculatus, and at which level
100% mortality occurred, within 76-hours and 50% mortality within 48
hours.

          Burdick and Lipschuety (1948) showed that potassium ferro- and
ferricyanide solutions in concentrations as low as 2 ppr. ar<. toxic to
fish life i°. the presence of sunlight.


Downing, Kathleen M.  1954.  The influence of dissolved oxygen concentra-
     tions on the toxicity of potassium cyanide to rainbow trout.  Journal
     Exp. Biol., 31:161-164.

          Downing (1954) found survival times of rainbc./ trout in concentra-
tions- of pot?csium cyanide in the range- of 0.105-0.155 ppm cyanide increased
with an increase in dissolved oxygen concentration between '10% and 100%
of air saturation.value, the effect being most marked with the lowest
concentration of cyanide.

          Downing (1954) also determined the rate of increase of survival
time with increasing concentration of oxygen did not appear to fall off
as air saturation value was approached.


Cairns, John, Jr., Arthur L. Buikemd, Jr., Alan G. Heath,, and Bruce C. Parker.
     1978.  Effects of temperature on aquatic organism sensitivity to
     selected chemicals.  Virginia Water Resources Research Center,
     Virginia Polytechnic Institute.  Bull.  106-112:24-71.

          Cairns et al.  (1978) determined  the median lethal concentration
of cyanide  (CN)  to goldfish at 5°C,  15°C,  and 30°C to be 3.25 +  1.95,
0..44,. and 0.28 ppm, respectively.

          Cairns et al.  (1978) determined  the median lethal.concentration
of cyanide  (CN)  to golden shiners  (Notemigonus crysoleucas) at 5°C, 15°C,
and 30°C to be 0.54 + 0.19, 0.31 + 0.06, and 0.30 + 0.08 ppm, respectively.

          Cairns et al.  (1978) determined  the median lethal concentration
of cyanide  (CN)  to bluegill at 5°C,  15°C,  and 30°C to be 0..24, 0.16, and
0.19 +  0.04 ppm, respectively.        •  •

         .Cairns et al.  (1978) found  that  the toxirity 01 cyanide to
Nitocris, a river snail; Fhilodina,  a  rotifer; Daphnia pulex; and Daphnia
niagna increased  as temperature  increased but decreased to the oligochacte
Aeolosoma.

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PHYSIOLOGY

Ruby, Sylvia M.,  D. George Dixor,  and Gerard.Leduc.  1979.   Inhibition of
     spermatogenesis in rainbow trout during chronic cyanide poisoning.
     Arch. Environ. Contain.  Toxicol., 8:533-5<44.

          Ruby et al. (1979),  in studying testicular tissue of rainbow
trout, found that cyanide reduced  cell division by about. 13% at 0.01 ag/1
and 50% ..at 0.03-mg/1 HCX.

          Ruby et al. (1979).   An  additional 9.5% of abnormal mitotic
figures (multipolar spiudles), were observed at 0.01 mg/1 HCN.

          Ruby et al. (1979).   Tissue from the highest cyanide concentra-
tion (0.03 mg/1 HCN) revealed  extensive necrosis of developing germ cells
within their cysts.

          Total number of dividing spermatogonia experienced a gradual
decrease under the influence of cyanide; the progesssive sequence of cell
divisions through the various  stages of prophase, roetaphase, and anaphase
were seriously disturbed by cyanide.

     1.   Twenty-four percent  of total cells were  in mitosis in prophase
          in the control fish  as compared to 88% and 55% in trout exposed
          to 0.01 and 0.03 mg/1 HCN, respectively.  The high number of
          cells in prophase should not be. interpreted as a stimulating
          effect of cyanide, but a result .of blockage of mitosis beyond
          prophase causing cells in that stage to  accumulate

     -2.   Metaphase was drastically reduced by cyanide, decreasing-from ••
          22% in controls to 6% and 12% in trout exposed to 0.01 and 0.03
          mg/1 HCN, respectively.

     3.   Cells in anaphase represented 54% of mitotic cells among control
          fish but the anaphase stage was completely absent from testes
          of rainbow trout exposed to 0.01 and 0.03 mg/1 HCN.

          Ruby et al. (1979) found that cyanide decreased the number of
spermatogonia in all stages of mitosis wj.Lh the greatest reduction at  the
h'gher concentration tested (0.03 mg/1 HCN).  Dividing spermatogonia
failed to complete mitosis, thus showing that developing germ cells
within the  testes of fingerling rainbow trout are  highly sensitive to  the
poison, suggesting that cyanide acts as an antimitotic agent which in
general affects mitosis.
                                                        t
          Ruby et al. (1979) felt that damage or death of spermatogonia
due  to low  concentrations of HCN could result in a significant reduction
in the ability of  rainbow trout to reproduce.

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Doudoroff, Fcter.   1956.   Some experiments  on the toxicityof. complex.
     cyanides to fish.   Sew.  and Ind.  Wastes, 28:1020-1040.

          Doudoroff (1956).   The estimated  median tolerance  limits (TL )
of sodium cyanide  in soft water to P.  prooielas at 20?C was 0.25,  0.24,m
and 0.23 ppm as CN  for 24-,  48-, and  96-hour experiments, respectively.

          Doudoroff (1956) determined  the TL  of a mixed solution of
sodium cyanide (564 ppm as CN ) and zinc suliate (394 ppm as Zn)  diluted
with soft water at 20°C to be 0.20, 0.19, and. 0,18 ppm as-CN~ during 24-,
48-, and 96-hour experiments, respectively.

          Doudoroff (1956) determined  the TL  of a mixed solution of
sodium cyanide (439 ppm as CN" ) and cadmium sulfate (528 ppm as Cd)
diluted with soft  water at 20°C to be  0.23, 0.21, and 0.17 ppm cyanide as
CN  for 24-, 48-,  and 96-hour experiments,  respectively.

          Doudoroff (1956) determined  a mixed solution of sodium cyanide
(600 ppm as CN ) and nickelous sulfate (355 ppm as Ni) diluted with soft
water at 20°C to be toxic at  all test  concentrations above 0.25 ppm as
CN  and felt that  cyanide combined with nickel evidently was less toxic
than free cyanide.                                        >

          Doudoroff (1956) found that  fisn  could withstand more than
1,000 times as much cyanide combined with nickel at pH 8.0 than at pH
6.5.  A doubling of the hydrogen ion concentration or reduction of pH by
little more than 0.3-pH unit  can result in  more than a 10-fold increase
of the toxicity of a mixture  of NaCN-NiSO.,.


Leduc, Gerard.  1978.  Deleterious, effects  of cyanide on early-life
     s:ages of Atlantic salmon (Salmo  salar).  Journ. Fish.. Res.  Bd.
     Canada, 35:166-174.

          Leduc (1978) found  during incubation of Atlantic salmo.i eggs  in
water of 5.4°C that yolk conversion efficiency was reduced at 0.02 mg/1
HCN and higher.  After hatching, this  trend was reversed.

          Leduc (1978) determinec1 the  sensitivity of Atlantic saTmon to
cyanide seems to stem from permanent damage caused early in morphogenesis
and cell differentiation stages, effects manifested by mortality and
congenital defects at very low cyanide levels.
Wilde, C. E., Jr. and R. B. Crawford.  1966.  Cellular differentiation in
     the auamniota.  III.  Effects of actinomycin 0 and cyanide on the
     morphogenesis of fundulus.  Experimental Cell Research; 44:471-488.

          Wilde and Crawford (J966) found that incubation of; mummichog,
Fundulus heteroclitus eggs or embryos of all stages in 2 by 10 3 M NACN
reduced oxygen consumption to zero and that embryos in cyanide undergo

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normal development patterns during cleavage and hlastulation,  whereas
pnstblastular development is inhibited.   Upon relief of the cyanide
inhibition, the enbryos renewed their morphogenesis at a normal rate.
During cleavage and blastulation, cyanide caused a marked acceleration of
glycolysis and nad little effect upon the steady-state level of ATP.
Glycolysis was not accelerated in postblastular embryos incubated in
cyanide while the cyanide inhibitor caused a marked reduction of nhe
level of ATP.
Leduc, Gerard.  1978. • Deleterious effects of cyanide on early life
     stages of Atlantic salmon (Saimo salar).  Journ. Fish.  Res.  Bd.
     Canada, 35:166-174.

          Leduc (1978) determined that cyanide at concentrations  as low
as 0,01 mg/1 HCN produced teratogenic abnormalities in Atlantic salmon
(Salmo salar) embryos.  He also found Atlantic salmon eggs exposed to 0.1
mg/1 HCN became discolored but at 0.01 mg/1 HCN there was no difference
Jrora control eggs.
BEHAVIOR

Leduc, Gerard.  1978.  Deleterious effects of cyanide on early life
     stages of Atlantic salmon (Salmo salar) .   Journ. Fish. Res. Bd.
     Canada, 35:156-174
          Leduc  (1978) observed a- striking-di-f ferenee-oe-tween— c
fish (Atlantic sdlmon fry) end cyanide-exposed fry at 0.08 and 0.10 mg/1
HCN.  The controls would swim actively in swarm when disturbed but thf:
poisoned fry remained almost motionless at the bottom.
BIOCONCENTRATIONS

Murachi, Shiro, Nanba Kenji, and Yukio Takeuchi.  1978.  Relation between
     the concentration of cyanide ion detected in carp and that in environ-
     mental water.  Fac. Fish. Animal Husb., Hiroshima Univ., Fukuyama,
     Japan.   In:  Chem. Abstracts, 91:'04.

          Huracl.i et al. (1978).  Relation between the concentration of
cyanide ion detected in carp and that in environmental water.  Hurachi, Shiro;
Nanba, Kenji; Takeuchi, Yukio (Fac. Fish. Anin. Husb., Hiroshima Univ.,
Fukuyama, Japan).  Hiroshima Diagaku Suichikusangakubu Kiyo  1978, 17(2.),
199-206 (Japan).  The toxicity of cyanide (I) to carp increased with a
rise in HjO temperature.  The concentration of I in Ihe blood was higher
than that in  the hepatopancreas and spleen, or the digestive tract of
carp exposed  to 10 ppm I solution for five hours.  L concentrations in
the skin and  muscle were higher than that, in the viscera of  carp which
had died in 10 ppm I solution.  Thus, I penetrated into the  skin and
through the gill.  I was detectable in the fish washed by rjnning water
for 24 hours  after death in 5 ppm I solution and also in the fish exposed
to  10 ppm I solution for 70 minutes after death from suffocation.
                                     9

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                                INVERTEBRATES
ACUTE EFFECTS

Taturn, William R.  1?68.  Field observations on the use of sodium cyanide
     in stream surveys.  Tennessee Game and Fish Commission 2^nd Annual
     Meeting of Southern Division of the American fisheries Society,
     Oct. 21-23, 1968.

        ••  Tatum (1968.) founu that sodium cyanide reduced the total benthos
population by about 77% when applied to a small mountain stream to collect
fish.
Cairns, John, Jr., Arthur L. .Buikema, Jr., Alan G. Heath, and Bruce C. Parker.
     1978.  Effects of temperature on aquatic organism sensitivity to
     selected chemicals.  Virginia Water Resources Research Center,
     Virginia Polytechnic Institute.  3ull. 106-112:24-71.
          Cairns et al. (1978) determined the 24-hour LCw; of KCN- to
Nitrocris <;p. at 5.0°C, 10.0°C, i5.0°C, 20.0°C, and 25.0°C to be 14.0,
13.0,  ll.fi,  1C.O, and 7.6 ppm, respectively.

        !.  Caiins et al. (1978) determined the 24-hour LC50 of KCN to the
rocifer Philodina acu .icornis to be 0.50 to 250.0 ppm at a temperature of
20.0°C.

        :  Cairns -et-al. (1978) determined fhe 24-hour LC50 of KCN to
Daphnia pulex at a water temperature of 5.0°C, 1C.O°C, 15.0°C, 20.0°C,
and 25.0°C to be 0.42, 0.33, 0.32, 0.15, and 0.003 ppm, respectively.

        .;.  Cairns et al. (197?) determined the 24-hour LC50 of KCN to
Aeolosoma headleyi, an annelid, at a temperature of 5°C, 10°C, 15°C,
20°C,  and 25°C to be 11.0,  100.0, 120.0, 160.0, and 160.0 ppm, respectively.

        .  C«:-'.rns el al. (1978) determined the 48-hour LCSO of KCN to
Nitocris sp. at  temperatures of 5°C, 10' C,  15CC, 20°C, and 25°C to be
13.6,  12.8,  10.0, 8.0, and  7.0 ppm, respectively.

        :  Cairns et al . (1978) determined the 48-hour LC50 of KCN to
Philodina acuticornis at 20°C to be 20.0 to 145.0. ppm.

        ;  Cairns et al. (1978) determined the 48-hour LCSO of KCN to
Daphnia pulex. at 5°C,  10°C, 15°C, 20°C, and 25°C t.o be 0.33, C.33, 0.18,
0.11,  ai;J O.CQ1  ppr^, respectively.

        ..  Cjirris et «1. (1978) determined the 48-hour LC5o of KCN to
Aeolosoma headleyi at 5°C,  10°C, 15°C,  20°C, and 25°C to be 10.0, 9.0, .
120.0,  160.0, and  160.0 ppm, respectively.
                                    10

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BORON
11

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                                    FISH
ACUTE AND CHRONIC

Leclerc, E.   1960.   The self-purification of streams and the  relationship
     betweta chemical and biological tests.  In:  Proc. 2nd Syrnp.  on
     Treatment of Va'.te Waters, P.C.G. Isaac 'ed.), pp. 281-316,  Pergamon
     Press,  London.

          Leclerc (l!*i/0) found -mininiUTr and maximum concentrations  of
3A5.0 mg/1 and 380.0 mg/1, respectively, of NajI^Oy to be tne  incipient
lethal level to fish in distilled water with A '..inimum temr-eratuie ot
19.0°C.
    ust^r,  J.  A.   19L6.  The -oxicity of certain weed killers  to  trout.
     Proc.  3rd Brit.  Weed Contr. Conf., 2:807-808.

          Alabaster (1956) determined the LC50 for Salmo gairdi. «ri  to be
392.0 mg/1  Na 26407 during a 48-hour pericl with a minimum  temr'.-iatuie of
18.0°C.  During a 24-hour period the I.Cso for Salmo gairdnevi  was determined
to be -S'0.0 mg/1  f^B^y, with a ruinimum water temperature ot   ">.Q°C.
WaUen. I.  E.. W. C. lireer, and R. Lar.ate- .  1957.  Toxicity  to  Gambusia
     aff inis of certain pure ch. .Tiicals iu  turbid waters.  Sewage Inu.
     Wa-tes, 29:69:.

          Wallen tt al. (1957) foui.d a 96-hour LC50 of  785.0  mg/1 of
Na2B407 for Gambusia aff inis .in tu. .bid water.
Leclerc, E.  1960.  T.ie self-purification of streams  a.^d-the  relationship..
     between chemical and biological tests.  In:  Pro*.  -!nd Symp.  on
     Treatment of W^ste Waters, P.C.G. Issac (ed.), pp. 281-316,  Pergamon
     Press, London.

          Leclerc (1960) found minimum and Maximum :' icipifn-t  lethal  level
concentrations of Na2B407 to fish to be 805.0 mg/1 and 870.0  rag/1,  respectively
in bard water with a minimun temperature of 17.0°C.


Wurtz, A.  1945. ..The action cf boric acid on crrtain  fish:'  trout,
     roach, rudd.  Ann. Sta. Bent. Hydrobiol. Appi. 1, 179; Water Pollut.
     Res., 20:1653(1947).

          Wurtz 0945) observed skin darkening on Salmo gairdneri  when
exposed to a concentration of" 905.0 mg/1 'of HBOs in water  with  minimum
and maximum temperatures of 14.0°C and 15.0°C, respectiv-.ly .
                                    12

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Wallen, I. E., W. C. Grter, and R. Lasater.   1957.   Toxicity to Gambusia
     at finis  of certain pure rhemicals in turbid waters.  Sewage Ind.
     Wastes,  29:695.

          Wallen et al. (1957) fou.id the 96-hour(LC5o of HB03 for Gambusia
affinis to be 1,014.0 mg/1 in turbid water.


Wurtz, A.  1945.  The action of boric acid on certain fish:  trout,
     roach,  rudd.  Ann. Sta. Bent. Hydrobiol  Appl. 1, 179; Water Pollut.
     Res., 20:1653(1947).

          Wurt= (1945) deterained the lethal concentration of HB03 to
Scardinus erythrophthaltnus during an 18-hour period to be 1,130.0 mg/1 in
water with irinimum and maximum temperatures of !4.0°C and 15.0°C, respec-
tively.  He  also found'the lethal concentration of HBOs to Hesperoleucus
during a 40-hour period to be 1,130.0 mg/^ in water of the same temperature
as noted above.


Wallen, I. E., W. C. Greer, and R. Lasater.   1957.   Toxicity to-Gambusia
     affinis  of certain pure chemicals in turbid waters.  Sewage Ind.
     W;istes,  29:695.

          Wallen et al. (1957) found the 48-hour LC5o of Na2B407 to
Gambusia affinis to be  1,790.0 mg/1 in turbid water.  The 48-hour LC5o of
HB03 to G. affinis was  1,900.0 mg/1 in turbid water-


Turnbull, H., J. G. DeMann; and R. F. Weston.  1954.  Toxicity of various
     refinery materials to freshwater fish.  Syrap.  on Waste Disposal  in
     the Petrol. Ind.,  Ind. Eng.  Chem., 46:324.

          Turnbull et al.  (1954)  determined the 24-hour LC50 of BF3  to
Lepomis macrochirus to  be  2,380.0 mg/1, with a minimum water.temperature
of 20.0°C~;
Wallen,  I.  E.,  W.  C.  Greer,  and R. Lasater.  1957.  Toxicity to Gambusia
      affinis  of certain pure chemicals in turbid waters.  Sewage  Ind.
      Wastes,  29:695.

           Wallen et  al. (1957) found~the 24-hour -LCSO of- NajB^O?  to
Gambusia affinis to  be  2,615.0 mg/1 in turbid water.        .
 Leclerc,  E.   1960.   The self-purification of streams and the. relationship
      between chemical  and  biological tests.  In:  Proc.. 2nd Symp-. on.
      Treatment of Waste Waters, P.C.G.  Issac (ed.), pp. 281-316, Pergamon
      Press,  London.
                                    13

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          Leclerc (1960) found the minimum and maximum incipient lethal
levels of HB03 to fish in water with a minimum temperature of 20.0°C to
be 3,260.0 and 3,530.0 mg/1, respectively.


Wallen, I. E., W. C. Greer, and R. Lasater.  1957.  Toxicity to Cambusia
     afflrii£ of certain pure chemicals in turbid waters.  Sewage Ind.
     Wastes, 29:695.

          Wallen et al. (1957) found the 24-hour LC50 of HB03 to Gambusia
affinis in turbid water to be 3,260.0 mg/1.


Wurtz. A.  1945.  The action of boric acid r>n certain fish:  trout,
     roach, rudd.  Ann. Sta. Bent. Hydrobiol. Appl. 1, 179; Water Pollut.
     Res., 20:1653(1947).
          Wurtz (1945)  found a concentration of 3,620.0 mg/1 of
caused behavioral distress and a concentration of 5,430.0 mg/1
caused equilibrium loss  in Salmo gairdneri in water with minimum and
maximum temperatures of  i4.0°C and  15.0CC, respectively.


(From EPA Redbook)

          Minimum lethal dose for minnows exposed to boric acid at 20°C
for six hours  uported  to be 18,000 to  19,000 mg/1 in distilled -water and.
19,000 to 19,500 mg/1 in hard water (Leclerc and Devlaminck, 1955; Leclerc,
1960).

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                                INVERTEBRATES
ACUTK

Anderson, B.  G.   1946.   The toxicity thresholds of varioi-s sodium salts
     determined  by the  use of Daphnia magna.   Sewage Works Journal,
     !8(l):82-87.

          Anderson (1946) found the incipient lethal level of NAB03 to
Daphnia magna to be 0.69 mg/1 during a 48-hour exposure time in Lake Erie
water,'with a water temperature of 25.0°C.


Anonymous.  1950.   Ohio River'Valley Water Sanitation Commission, Subcommittee
     on Toxicities, Metal Finishing Industries Action Committee.  Report
     No. 3.

      '  •  Anonymous (1950) found the incipient lethal level of N^BjO? to
Daphnia magna to be 26.0 mg/1.


Jones, J.R.E.  1941.  A study of the relative toxicity of anions with
     Polycelis nigra as test animal.  J. Exptl. Biol., 18:170-181.
      f
          Jones  (1941)  found the incipient lethal lev°l of Na2B407 H3B04
to Polycelis nigra during a 48-hour period with a minimum temperature of
14.0°C and~a maximum temperature of 18.0°C to be 280.0 mg/1.
                                    15

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                              AQUATIC  PLANTS
Stanley, R. A.   1974.   Toxicity  of  heavy metals  and salts .to Eurasian
     watennilfoil (Myriophyllum  spicaturo L.) Arch.  Environ.  Contain.
     foxicol.,  2:331-341.

          Stanley (1974) found that 40.30 mg/1 cf  B407 caused 50% root
inhibition in Myriophyllum spicatum during  a 32-day .exposure period in
tap water at a water temperature of 20.0°C.
                               MICROORGANISMS
Sheets, W. D.  1957.  Toxicity studies  of  metal-finishing wastes.  Sewage
     Ind. Wastes, 29:1380.

          Sheets (1957) observed  a  50%  reduction of biological oxygen
demand in sewage organisms  exposed  to 87.0 mg/1  of HBO3 in buffered
water.
                                    16

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SULFIDE
17

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                                    FISH

                             (ACUTE AND CHRONIC)
EGGS AND HATCHING
Smith, Lloyd L., Jr., Donavon M. Oseid, Gary L. KimbalJ., and Sayed M.
     El-Kandelgy.  1976(a).  Toxicity of hydrogen sulfide to various life
     history stages of bluegill (Lepomis macrochirus).  Trans. Am. Fish.
     Soc., 3:442-449.

          Smith et al. (1976a) in acute tests found that newly fertilized
eggs tested at 5.9 aad 6.1 mg/1 02 "and 21.9°C hatched in 77 and 66 hours,
respectively, with a mean LCso'of 0.0218 mg/1 H2S.

          Smith et al. (1976a) in two chronic tests with durations of 130
and 316 days started with newly fertilized bluegill eggs hatched in H2S
concentrations which varied from 0.0018 to 0.0136 mg/1.  Survival decreased
to 14% and 11%, with concentration of H2S increased to 0.0136 and 0.0127
mg/1, respectively.


Adelman, Ira R. and Lloyd L. Smith, Jr.  1970.  Effect of hydrogen sulfide
     on northern pike eggs and sac fry.  Trans. Amer. Fish. Soc., 3:501-509.

          Adelman and Smith (1970) conducted flow-throup.h bioassays on
northern pike eggs (Esox lucius L.) to test the effect of H2S at two
concentrations of oxygen.  At an oxygen concentration of approximately 2
ppm mean median tolerance limits (TL ) for eggs were 074117 "0'.0'76~0'.~038;""
0.034, and 0.030 ppm H2S for 24, 4B,m72, arid 96 hours, and for the duration
of the embryonic period, respectively.  At an oxygen concentration of
approximately 6 ppm median-tolerance limits (TL ) were 0.181, 0.046,
0.041, 0.037, and 0.032 ppm H2S for 24, 48, 72,mand 96 hours, and for the
duration of the embryonic perioc?*, respectively.  The pH for each test
varied from 7.3-8.0.             .

          Eggs subjected to H2S resulted in increasing percentages of sac
fr> vjrh anatomical malformations.  At the highest K2S concentration,
0.061 ppm, 28% of hatched fry were abnormal.  Eggs hatched at the lowest
H2S concentrations, 0.004 ppm, produced larger sac fry than controls.
Maximum possible safe level of H2S for egg-; is between 0.014 and 0.018
ppm.                                                  .

          Adelman and Smith (1970) determined that H2S and oxygen acted
independently  in causing mortality with no significant interaction between
the two.
Colby, Peter J. and Lloyd L. Smith,  Jr.   1967.  Survival of walleye eggs
     and  fry on paper  fiber  sludge deposits  in  Rainy River, Minnesota.
     Trans. Amer. Fish. Soc. 96(3):278-286.

                                     18                      . V

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          Colby and Smith (1967)  found  the  greatest walleye (Stizostcdion
vitrcum) egg mortality to be  over wood-fiber deposits where dissolved
sulfido concentrations reached  0.58  ppm within 2 cm of the bottom.   They
lelt that dissolved oxygen, HaS,  or  C02 nay singularly or in combination
account for hi^h egg mortalities  over  fiber deposits.  Eggs and sac fry
died within two days when plated  on  the bottom where the dissolved sulfide
concentration was 0.28 ppm.   They found walleye eg£S are more tolerant of
dissolved su'fide when ample  dissolved  oxygen is present and that dissolved
sulfidc concentrations of 0.33  and above were'toxic to eyed eggs and
newly hatched fry.
SAC FRY

Adelman, Ira R. and Lloyd L.  Smith,  Jr.   1970.•  Effect of hydrogen sulfide
     on northern pike eggs and  s'ac  fry.   Trans.  Amer.  Fish. Soc., 3:501-509.

          Adelman and .Smith (1970)  conducted bioassays on northern pike
(Esox lucius L.) sac fry to test  the effect of HgS at two concentrations
of oxygen.  The pH varied from  7.3-8.0.   Median TL  values for sac fry
were 0.035, 0.016, 0.012, and 0.009 ppm  H2S at 2 ppm oxygen and 0.16,
0.047, 0.030, and 0.026 ppm H2S at  6 ppra oxygen for 24, 48, 72, and 96
hours, respectively.  Sac fry hatched from eggs held at the higher l^S
concentrations were smaller than  the controls.   S?c fry subjected' to H2S
showed decreased growth rates at  the higher concentrations.  Maximum
possible safe level of H2S for  sac  fry is between 0.004 and 0.006 pp. for
96-hour exposure.  They also found  a significant interaction between
reduced dissolved oxyger and ^S  in causing mortality of sac fry.


Colby, Peter J. £-•: Lloyd L.  Smith,  Jr;   1967.   Survival of walleye *ggs
     and fry on-paper fiber sludge  df osits in Rainy River, Minnesota.
     Trans. Amer. Fisu. ioc., 96(3):278-286.

          Colby and Smith (1967)  determined walleye sac fry are more
vulnerable to dissolved sulfide than eggs.
Smith, Lloyd L. , Jr., Donavon tf.  Oseid,  Gary L. Kiirball, and Sayed M.
     El-Kandelgy.  1976(a).   To.xicity of hyd.o~°n sulfide to various life
     history stages of bluegili (Lepomis macroc.iirus) .   Trans. Am. Fish.
     Soc., 3:442-449.

          Smith et al. (1976a) found that sac fry tested at 21.7°C ar.d
5.8 mg/1 02, four days after egg fertilization, had a 96-hour LCso of
greater than 0.0435 nsg/1 H2S and at nine days was 0.0253 mg/1
          Smith et al. (1976a) determined that advanced feeding fry
tested at 21.8°C and 6.0 mg/1 Oa,  39 days after egg fertilization, had an
LC50 at 96 hours of 0.0131 mg/1 and after 8 days 0.0128 rag/1 HjS.
                                   19

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JUVENILE-ADULT

Hayden, E. P., H. R. Amberg, and R.  E.  Diraick.   1952.   The  effect  of
     Kraft irill waste components on certain saltnonoid  fishes  of  the
     Pacific Northwest.   Tappi, 35(12)-.545-549.

          Hayden et al.  (1952) conducted static  bioassays  concerning the
effects of hydrogen sulfide and sodium  sulfide  on king salmon (Oncorhynchus
tshawytscha), silver salmon (0. kisutch),  and coa: tal  cutthroat  trout
(Salmo clarkii clarkii).   Temperatures  during tests en king salmon were
17.5°C + 2°, those with silver salmon were 15°C  + 3°,  and with coastal
cutthroat trout were 12°C + 3°; DO levels  were  from 9-12 ppm. The minimum
lethal concentration of hydrogen sulfide for king salmon and  cutthroat
trout was 1.0 ppm and 1.2 ppm for silver salmon.  He found  the minimum
lethal concentration of sodium sulfide  for king  salmon to  be  3..5 ppm, for
silver salmon 3.1-ppm, and cutthroat trout 3.0  ppm.
Wallen, I. E., W. C. Greer, and R. Lasater.   1957.   Toxicity to Oambusia
     affinis of certain pure chemicals in turbid waters.   Sewage Ind.
     Wastes, 29:695.

          Wallen et al. (1957) found the 24-, 48-,  and 96-hour median
tclerance limits of sodium sulfide to Gambusia affinis to be 750 ppm,  750
ppm, and 750 ppm, respectively, with a water temperature  of 21-25°C and a
pH of 7.6-11.6. .              .

          Wallen et al. (1957) fcund the-24--_,-48--,-and_96rhour median
tolerance limit ,of sodium sulfite to Gambusia affinis to  be 2,600,  2,600,
and 2,600 pi-m, respectively, with a-water temperature Oi  18-26°C and a pH
of 7.1-7.9.

          Wallen et al. (1957) found the 96-hour TL  of ferrous -sulf.ide
to G. affinis to be greater than 10,000 ppm at a temperature of 20-26°C
and pH of 1.1 to 8.4.

          Wallen et al. (1957) found the 96-hour TL  of ferrous sulfitc
to G. affinis to be 350 ppm at a temperature of 20-2l°C and pH of 3.2-6.9,

          Wallen et ul. (1957) found the TL  of ammonium sulfide to
Gambusia affinis to be 248 ppm at a water temperature of 21°C and pH of
7.0-8.8.

          Wallen et al. (1957) found the TL  of ammonium sulfits to
Gambusia affinis to be 240 ppm at a water temperature of 20-2l°C and pH
of 2.7-6.7.
Smith, Lloyd L.,!Jr. and Donavon M. Oseid.  1975.  Chronic effects of low
     levels of hydrogen sulfide on freshwater fish.  Progress in Water
     Technology,; 7(3/4):599-605.
                                    20

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          Smith and Oseid (1975).   Fathead  minnow eggs,  temperature 24°C,
96-hour LCGO of 0.0350 rag/1  H2S,  and  a  threshold l.Cso of 0.0351  mg/J  112S
at 4-8 days.


Chronic Tests - Fish-Bluegills

Smith, Lloyd L. , Jr. and Donavon  M. Oseid.  '1975.,  Chronic effects of low
     levels of hydrogen sulfide on freshwater fish.   Progress  in Water
     Technology, 7(3/4):599-605.

          Smith and Oseid (1975).  . Young-of-year bluegills after 826 days
of exposure as young-of-year to 0.0070  mg/1 H2S, their mean weight was
approximately 63% of controls.

          Smith and Oseid (1975).   After 97 days exposure of fish started
as. adults to levels of hydrogen sulfide up  to 0.0078 mg/1 there  was no
appreciable difference from controls  except at 0..0014 mg/1 where there
was a significant increase in growth.

          Smith and Oseid (1975).   Reproduction (number of eggs  per gram
of female) was significantly reduced  at 0.0007 mg/1 H^S and completely
inhibited at 0.0027 and 0.0078 mg/1 H2S.

          Smith and Oseid M975).   In an experiment started with young-of-year
bluegill, no spawning occurred at concentrations of 0.0015 mg/1  and
higher.  This failure of egg deposition appeared to be due to inhibition
or absence of spawning behavior since apparently normal numbers  of viable
eggs were found in ovaries of nonspawning fish.


Smith, Lloyd L., Jr. and Donavon  M. Oseid.   1974.  Effect of hydrogen
     sulfide on development and survival of eight- freshwater fish species.
     In:  Early Life History of Fish, J.H.S.  Blaxter (ed.), 415-430.
     Springer-Verlag, New York, NY.

          Smith and Oseid (1974).   Fathead .minnows exposed for two repro-
ductive cycles at levels of H2S between 0.0004 and 0.0069 rag/1 and at
23°C during the first cycle had no significant difference in the number
of eggs laid per female.  In the  second cycle, a slight reduction in eggs
per female occurred with 912 in a control and 791 at O.G069 mg/1 H2S.
Survival of spawning adults was lower in both Cycles at the highest
concentration  (0.0069).

          Smith and Oseid (1974).   Bluegills  .held for 826 days had no
reproduction at 0.0018 mg/1 and significantly reduced fingerling-to-adult
survival at 0.0073 mg/1.

          Smith and Oseid (1974).  Fale bluegills during a 46-day test at .
0.0014 mg/1 H2S showed a slight alteration in spawning  behavior, at
0.0027 mg/1 it was  greatly reduced, and at 0.0078 mg/1  none occurred.
                                    21

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          Smith and Oseid (1974).  Brook trout held for 37 days prior to
spawning (total exposure 76 days) at levels of 0.0055-0.0128 mg/l 1I2S
deposited fewer eggs per female tha^. controls.  At 0.0128 rag/1 H^S, the
number of eggs laid was 30% of controls and at 0.0055 mg/l was 46%.  No
apparent effect on behavior was noted at 0.0079 mg/l h^S, but at higher
concentrations spawning activity was greatly reduced.

          Saith and Oseid (19/4).  Walleye, fathead minnows, bluegill,
r.iinbow trout, brook trout, northern pike, and goldfish eggs were sub-
jected to various levels of H2S from O.OC5 to 0.059 mg/l to determine
effects on incubation.                    :

          Smith and Oseid (1974).  In egg tests on walleye, percentage
hatch dropped from 86% of control at 0.012 mg/l to 9% at 0.159 mg/l, and
hatching time was extended from 22 days in the control to 26-27 days at
all levels of H2S treatment.

          Smith and Oseid (1974).  Survival of eggs incubated at 0.016-0.053
mg/l dropped from 87% at 0.016 mg/l to 17% at 0.058 mg/l.  Hatching was
extended from 6 dayc in t!;e control to 8-9 days at the higher h^S concentrations.

          Smith and Oseid (1974).  Eggs tested at 0.011-0.035 mg/l survived
at 63% of controls in the lowest concentration and 13% in the highest.
Hatching time did not vary significantly at the various levels.

          Smith and Oseid (1974).  Eggs incubated at 0.005-0.01! mg/l KZS
varied from 67% at 0.005 mg/l to 50% at 0.011 mg/l.  Brook trout eggs
laid by adults held in the same test conditions during maturation and egg
deposition had much poorer survival with 45% at 0.007 mg/l and no survival
at 0.011 rag/1.

          Smith and Oseid (1974).  Eggs incubated at 0.006-0.047 mg/l
survived at the rate of 76% in the lowest to 4% in the highest concentration.

          Smith and Oseid (1974).  Eggs incubated at 0.018-0.058 mg/l
varied from 88% of controls ct the lowest level of HaS to 6% at the
highest.                                                  :

          Smith and Oseid (1974).  Eggs held at 0.010-0.029 trg/1 hatched
at the rate of 95% in the lowest level and 12% in the highest.


Smith, Lloyd L., Jr., D. M. Oseid, and L. M. Olson.  1976(b).  Acute and
     chronic toxicity of hydrogen sulfide to the  fathead minnow,' Pimephales
     promelas.  Environ. Sci. .echnol;, 10(6):565-568.

          Smith et al. (1976b).  Acute toxicity tests of fathead minnows
and eggs to hydrogen sulfide.

          Smith et al. (1976h).  Six bioassays were conducted with' fa.the.ad
minnow eggs at temperatures from 23.8-24.2°C, oxygen from 5.6-6.0. mg/l,
                                   22

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and pli of 7.9 to determine the toxicity of hydrogen sulfide.   The 96-hour
LCSO varied between tests froai 0.0291-0.0933 mg/1,  with a mean of 0.0536
mg/1 H2S.  The LCso for the incubation period which ranged from 5-8 days
had a mean of 0.0258 mg/1 H2S.

         . Smith et al.  (1976b).   Tests on fry started within 24 hours of
hatching were run at 24°C and 5.4-6.2 mg/1 02.  The meao LC5o varied from
O.C208 mg/1 at 24 hours to 0.0107 mg/1 H2S at 96 hours.  Mean LTC at six
days was 0.0093 mg/1 H£S.  LCSO at 96 hours varied  from 0.0100-0.0115
mg/1 H2S and LTC from 0.0087-0.0101 rag/1..H2S.

          Smith et al.  (1976b).   Bioassays were performed on juveniles at
six temperatures:  6.5°C, 7.6<5C, 10.0°C, 15.0°C, 20.2°C, and 25.0°C.
LC50 at 96 hours ranged from 0.7754 mg/1 at 6.5°C to 0.0423 mg/1 H2S at
25.0°C.  The ..cute toxicity of H2S to juvenile fish was greatly changed
by temperature.

          In chronic exposure to H2S from egg through two generation? of
laboratory-cultured fathead minnows in flow-through bioassays, adverse
effects on growth, survival, and fecundity occurred above 0.004 mg/1 H2S.
Chronic exposure of wild stock up to 346 days caused adverse effects on
growth and survival above 0.008 mg/1 H2S.

          At comparable temperatures, apparent.safe levels with long
exposure were 5- to 7-fold lower than the 96-hour LC$Q for both stocks.
Oseid, Donavon M. and Lloyd L.  Smith,  Jr.   1972.   Swimming endurance and
     resistance to copper and mulathion of bluegills treated by long-term
     exposure to sublethal levels of hydrogen sulfide.  Trans. Amer.
     Fish. Soc., 4:620-625.

          Oseid and Smith (1972) found that exposure of bluegill to H2S
increased their resistance to copper.   Bluegills  conditioned at the
lowest H2S treatment (0.0004 mg/1) had a significantly longer survival
time when treated with malathion than controls, but at higher levels of
H2S there was no increased resistance to malathion.
Shelford, Victor E.  1917.  An experimental study of the effects of gas
     waste upon fishes, with especial reference to stream pollution.
     Bull. Illinois State Laboratory Nat. History, Vol. XI,.Article..VI,
     pp. 395-398.

          Shelford (1917) foi-.n-J that carbon bisulfide, 0.05 cm per liter
of water', intoxicated fish but. the fish recovered after 1-1/2 hours in a
test in a closed bottle.
                                   23

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Smith, Lloyd L.,  Jr., Donavon  M.  Oseid, Gary  L.  Kimball,  and  Say<»d M.
   .  El-Kandelgy.  1976(a).  Toxicity  of  hydrogen  sulfide to  various life
     history stages of bluegill  (Lepoais  macrochirus).  Trans.  Am. Hsh.
     Soc., 3:442-449.

          Smith et al- (1976a) felt  that  monitoring programs  may not be
able to detect the lovest levels  of  h^S shown to have  toxic influence in
the laboratory and that any detectable concentration in the field should
be considered detrimental to fish production.
Huckabee, John W., C. Phillip Goodyear,  and  Ronald D.  Jones.   1975.   Acia
     rock in the Great Smokies:   Unanticipated impact  on aquatic biota.of
     road construction in regions of sulfide mineralization.   Trans.  Am.
     Fish. Soc., 4:677-684.  -

          Huckabee et al. (1975)  found  that  leaching of sulfide-rich
rocks in a mountain stream was responsible for trout and salamander
mortalities causing levered  pH to 4.D and sulfate concentrations as high
as 56.0 mg/1; water teeperature varied.
Darnell, Reyneat M., Willis E.  Pequegnat,  Bela M.  James,  Fred J. Benson,
     and Richard A. Dafenbaugh.   1976.   Impacts of construction activities
     in wetlands of tbe United  States.   U.S.  EPA Corvallis Environmental
     Research Laboratory, Corvallis,  Oregon.   Contract No. 68-01-2452,
     p. 267.

          Darnell et al. (1976)  mentioned  that construction activities
increased hydrogen sulfide'levels by increasing sedimentation and burial
of. organic matter, levering of  the oxygen  and the pH of the water, digging
and stirring up of bottom sediments,  low-level release of water fronr
dams, canalization of coastal marshes,  reduction of minimal flow rates
and internal circulation patterns, and  introduction of sulfides.and..acids
into wetland environments.  He  (1976) stated  that hydrogen sulfide enters
aquatic systems from the sediments and  eggs and young of fishes and many
of the important fish food organisms are apt  to be subject to the highest
concentrations when stagnant conditions develop.
Adelman,  Ira R. and Lloyd L. Smith, Jr.   1972.   Toxicity cf hydrogen
     sulfide to goldfish (Carassius auratus) as influenced by temperature,
     oxygen, and bioassay techniques.  Journ. Fish...Res. Bd—Canada,
     29(9):1309-1317.

          Adelman and Ssith (1972) determined the mean 96-hour TLso of
hydrogen  sulfide to goldfish to be 530 |Jg/l at 6.5°C to 44 |jg/l at 25°C.
The  log of  the TL5Q increased proportionally to a decrease in the log of
temperature.

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          Adelman and Smith (1972) conducted acute bioassays with hydrogen
sulfide at various oxygen concentrations with goldfish.  In bioassays
without acclimation, the mean TL5o was 71 |Jg/l HgS at 6.0 tng/1 oxygen and
53 (Jg/1 HjS at 1.5 mg/1 oxygen.  In tests with acclimation, *.he siean
TLs0s were 62 and 48 yg/1 H2S at the same oxygen concentrations.  In 8 of
10 pairs of'tests, the bioassay conducted at the lower oxygen concentration
resulted in a lower 96-hour TLso-  However, the rate of increased toxicity
with lowered oxygen is not very great, with the 9C-hour TL5o reduced only
about 26% between oxygen concentrations of 6 and 1 mg/1.  Prior acclimation
of goldfish to the oxygen concentration of the bioassay had little effect
on "acute H2S toxicity except at very low oxygen concentrations.


Chevalier, J. R.  1973.  Tox.icity of sodium sulfide to common shiners -
     dynamic bioassay,  Tappi, 56(5):135-136.

          Chevalier (1973) exposed connon shiners  ^otropis cornutus
Hit-hill) to sodium sulfide in concentrations from 0.33 to 3.95 ppm using
a dynamic bioassay.  Median tolerance Limits (TLso) were calculated for
24, 48, ami 96 hours; results were 2.(.3, 1.72, and 1.64 ppra, respectively,
with a water temperature between 13.0JC and 14.0°C.  Most mortality
occurred within the first 48 hours
Smith, Lloyd L., Jr., Donavon M. Oseid, Gary  L. Kiniball, and Sayed M.
     El-Kandelgy.   1976(a).  Toxicity of hydrogen  sulfide to various life
     history stages  of bluegill  (Lepomis macrochirus).  Trans.-Am... Fish.
     Soc., 3:442-449.

          Smith et  al.  (1976a) ran  five tests for  96  hours on juvenile
bluegill with  a mean size  of 3.9 cm; temperatures  varied from 20.1°C to
19.9°C with 02  ranging  from 5.7  to  6.6 mg/1.   The  mean 96-hour LC50 was
0.0478 mg/i H2S.  The LC50 of one t^st run  frr 8 days to threshold was
0.0491 rag/1 H2S and a second run-for.10. days  w?s.0.0468 mg/1.

          Smith et  al.  (1976a) ran  seven acute tests  on adult bluegill
held for 31-174 days prior -o bioassay.  Mean length  of fish was  12.1 cm,
temperatures were  19.6°C to 20.3°C,  and 02  levels  weie 4.6 mg/1 in one
test and in otheis  ranged  from 5.8  to 6.4 mg/1.  The  mean 96-hour LCgc
was 0.0448 mg/1 and showed no relation to 02  levels.

     •     Smith et  al.  (1976a) tested juvenile bluegill with a H2S concentra-
tion of 0.0092 mg/1 and  resulted in 100% survival  at  717 days-and- 70% by
the 826th day.


Bonn, Edward W. and Billy J. Follis.  1967.   Effects  of hydrogen  sulfide
     on channel catfish, Ictalurus  punctatus.  Trans. Amer. Fish. Soc.,
     96:31-37.
                                    25

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          Bonn and Follis (1967) determined at a pH of 7.0 the TL  of
un-ionized hydrogen sulfide was found to be 1.0 ppm for fingerling chanac'1
catfish, 1.3 for advanced fingerlings, and 1.4 I'or adult catfish.  This
indicates channel catfish have a greater resistance to I^S with an increase
in size or age.
Van Horn. Willis M.,  J. B. Anderson, and Max Katy.  1949.  The effect of
     Kraft pulp mill  wastes on some aquatic organisms.  Trans. Amer.
     Fish. Soc., 79:55-63.

          Van Horn, Anderson, and Katy (1949) determined the minimum
lethal concentration  of H2S to Lake Emerald (Notropis atherinoides) and
spotfin shiners (N. spilopterus) to be 1.0 ppm in a period of 120 hours;
temperature equals 18."OcC,'pH equals 7.6-7.8, and total alkalinity 140-160
ppm.  They determined the minimum lethal concentration'of sodiiun sulfide
to the same fish as above to be 3.0 ppm in a period of 120 hours with
same temperature, pH, and total alkalinity.
PHYSIOLOGY

Smith, Lloyd L. , Jr., Donavon M. Oseid, Gary L. Kimball, and Sayed M.
     El-Kandelgy.  19/6(a).  Toxicity of hydrogen sulfide to various life
     history stages of bluegill (Lepomis macrochirus)..  Traiis. Am. Fish.
     Soc., 3:442-449.

          Smith et al. (1976a) shewed that growth rate of blueg?.1! started
as eggs and maintained- for 316 days exposed to 0.0136 mg/1 H2S h?d a mean
weight of 40.8% of controls.  After-11'3-days-erf—exposure-lir-a—sec^nri—treslr
at 0.0127 mg/1, mean weight was 86.7% of control.

          Smith et al. (1976a) in tests started with juvenile bluegill
showed a retardation of growth at all H2S levels tested above 0.0022
mg/1.  At 0.0092 mg/1 H^S, the mean weight was approximately 35% of the
control after 392 days.

          Smith et al. (1976aJ found in tests with adult bluegills there
was no significant decrease in growth at concentrations less than 0.0107
mg/1 H2S.  The mean weight of fish held for 288 days at 0.0149 mg/1 was
59% of control, and in a test running 200 days fish at 0.0144 mg/1 H2£
weighed 73% of control.

          Smith et al. (1976a) found a significant decrease in food
intake (live minnows by immature bluegills) only above 0.0085 mg/1 H2S
during a 28-day period.  Conversion efficiency ranged from 3.57 in the
control to 1.27 at 0.0144 mg/1 H2S.

          Smith et al. (197u?) in.studies on bluegills found when growth
rate and long-term surv. .v.l were used as indices, fish started as eggs
were the most sensitive to H2S, whereas fish started as juveniles and
adults were less affected.

                                   26

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          Smith et al.  (1976a)  in  one  Lest  running for 826 days found
bhiogills failed to spawn  in  treatments  of  0.0022  to  0.0092 nig/1  H2S,
whereas control fish spawned  with  an average  of 9,928 eggs/female and 130
e8Ss/8 °f female.   lo a second  test, prespawning adults,  for a period of
97 days, produced  155.5 eggs/g  of  female in the -control,  100.8 eggs/g of
female at 0.0010 mg/1,  and 51.1  eggs/g of female at 0.0021 mg/1,  while a
concentration of 0.0041 mg/1  h^S resulted in  no egg deposition.

          Smith et al.  (1976a)  determined that a failure  to deposit eggs
appeared directly related  to  inhibition  of  spawning behavior and  that an
examination of the gonads  in  fish  which  did not spawn did not indicate
•any apparent malformation  or  lessening of -the- number- ot eggs.
Oseid, Donavon M. and Lloyd ,L.  Smith,  Jr.   1972.   Swimming endurance and
     resistance to copper and malathion  of bluegills treated by long-term
     exposure to sublethal levels  of hydrogen sulfide.   Trans.  Amer.
     Fish. Soc., 4:620-625.

          Oseid and Smith (1972)  found the gross  effects of long-term
exposure of bluegill to H2S were  reduced growth in the  highest  concentra-
tion (0.0146 mg/1) and progressively increased gill irrigation  rate with
increased concentrations of
          Oseid and Smith (1972)  found that YOY bluegill which had. been
exposed to higher concentrations  of H2S had a slightly increased capability
to endure swimr.ing stress at a  c'   °.ntration of 0.0004 mg/1 but that all
other swimming tests showed fis   •; hr adversely affected.
          Oseid and Smith (1972)  f  md that extended exposure to
levels of 0.-0015 ng/1 and greater .educed the physical capability of
bluegill.
Bonn, Edward W. and Billy J.  Follis.   1967.   Effects of hydrogen-sulfidc
     on channel catfish, Ictalurus punctatus.   Trans. Amer. Fish. Soc.,
     96:31-37.

          Bonn and Follis (1967) found" when channel catfish were exposed
to sublethal concentrations of un-ionized J^S (concentrations approaching
within 0.2 ppm of the TL  for a given pH) they exhibited nervousness and
excessive movement as if attempting to escape the poisonous gas.
Shelford, Victor E.  1917.  An experimental study of the effects of gas
     waste upon fishes, with especial reference to stream pollution.
     Bull. Illinois State Laboratory Nat. History, Vol. XI, Article VI,
     pp. 395-398.

          Shelford (1917) found that hydrogen sulfide is more toxic to
fish when oxygen is low.  He found 2 cc per liter fatal to fishes.  Water
                                     27

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exposed to the air with 4 cc per liter killed fishes in 18-24 hours.
Hardy species of fish  lived in 1-1.8 rr per liter without apparent
injury.
Terrans, Eugene Leslie.  1980.  Acute toxicity of hydrogen sulfide to
     fish during harvesting operations on commercial catfish farms:
     Cause, prevention, and cure.  Dissertation, University of Oklahoma,
     pp. 1-123.

          Terrans  (1980) found that hydrogen sul-f ide was a competitive
inhibitor of cytochrome oxidase in vitro in fathead minnow brain homogenate.
A 10 6 M sulfide concentration increased the K  of the enzymatic oxidation
of cytochrome C from  1.45 times, 10"5 M to 1.00mtimes 10"4 M.

          Terrans  (1980) found that hydrogen sulfide at very low concentra-
tions inhibited cytochrome oxidase in vitro.  The inhibition 2^ the
enzyme in a channel catfish brain pr.paration was 18% with 10 7 M J^S,
64% with 10"6 M, and  100% with lo""1 M.

          Terrans  (1980) f.'ound the effect of acute exposure to hydrogen
sulfide on the in  vivo inhibition of cytochrome oxidase varied with the
type of tissue tested.  At the point of respiratory arrest, the tissues
of the fathead minnow showed cytochrome oxidase activities ranging-frora
control levels in  the testes to a 55% inhibition in the kidney.  In the
channel catfish, the  effect ranged from a 28% decrease of the brain
cytochrome oxidase activity, to a_66% decrease in the heart whea fish were
sampled at the point  of respiratory arrest.

          Terrans  (1980) found the inhibition of 'cytochrome oxidase in
the channel catfish brain and gill was affected by the un-ionized sutfide
concentration.

          Terrans  (1980).  When fish were exposed-to a 0..1 mg/1 H2S at
10°C, the cytochrome  oxidase of the brain was not affected by even a
30-minute exposure.   The enzyme of the gill, however, was inhibited 15%
after only 5 minutes  and 39% after 30 minutes in the sulfide.

          Terrans  (1980) found that levels of blood lactate in fingerling
channel catfish rose  from 11.6 mg/100 ml to 38.1 mg/100 ml when exposed
to 0.1 mg/1 un-ionized sulfide at a temperature of 20°C.

          Terrans  (1980) found the inhibition of cytochrome. oxidase by
hydrogen sulfide is reverrible within a short period of time.  The enzyme
activity of channel catfish brain recovered from a 50% inhibition to
control levels within six hours in fresh water at 10°C and at a similar
rate at 20°C.  At  20°C, the recovery of the enzyme in both the brain and
the gill proceeded at the same rate.

          Terrans  (1980) found that raising the pH from 5.0 to 7.5 reduced
the un-ionized sulfide concentration by 84% and decreased the inhibition
                                    28

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of the en/yme in channel catfish by 20%.  Apparently any condition that
may lower the intramitochondrial pH, such as metabolic acidosis, will
increase the toxicity of sulfide.

          T»rrans (1980) determined the effect of sulfide is increased
greatly with increasing .temperature.  A concentration of 0.5 mg/1 I^S at
10°C was required to produce the same inhibition of enzyir.e activity in
vivo as 0.1 mg/1 F^S at 20°C in studies with channel catfish.

          Terrans (1980) found the uptake of snlfide ceased with the
onset of respiratory arrest and riote'l thot channel catfish that show
severe symptoms of sulfide poisoning during commerical harvesting can be
saved if they are removed to fresh water immediately.
                                    2o

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                                INVERTEBRATES
ACUTE AND CHRONIC

Anderson, B. G.   1946.   The  toxicity thresholds of various sodium salts
     determined  by the  use of  Daphnia mr.gna.   Sewage Works Journal,
     18(l):82-87.

          Anderson (1946) determined the  toxicity thresholds of several
so-'.ium salts to  Daphnia niagna.  The toxicity  of sodium sulfide was 9.4
ppm whenever the toxicity of sod\um sulfite was 440 ppra.


Oseid, Donavon M. and Lloyd  L.  Smith, Jr.   1974.  Chronic toxicity of
     hydrogen sulfide to Gamma r as pseudolimnaeus.  Trans. Araer. Fish.
     Soc., 4:819-822.

          Oseid  and Smith  (1974; determined  the mean 96-hour LC$o for
Gamma rus pseudolimnaeus as 0.022 mg/1 of  H2S.   They indicated that 0.002
mg/1 of HaS is the maximum safe concentration for Gamma rus.
          Oseid and Smith  (1974)  did  acute  tests on Gamma rus pseudolimnaeus
with hydrogen sulfide concentrations  ranging from 0.008 to 0,093 mg/1.
Temperature varied from 17.8-18.1°C,  oxygen from 5.8-7.4 uig/1, and pH
from 7.7-7.9.  Total alkalinity ranged  from 222-232 mg/1.  The total
length of organisms exclusive of  antennae were 0.7-1.2 cm.  For the
successive tests,, the 96-hour LCsos were  0.022, 0.022, 0.024, and 0.021
mg/1 H2S.  A single threshold test  run  through 18 days gave an. LCso value
of 0.011 mg/1 H2S.

          Oseid and Smith  (1974)  determined hydrogen sulfide concentration
in the four chronic tests  with Gamma rus pseudolimnaeus varied from 0.0007
to 0.0192 mg/1.  Temperatures in  three  tests varied from 17.1-17.8°C and
in one -test was 18-18. 2°C.   Dissolved oxygen ranged from-7.4-8.9-mg/l.
Survival at the higher levels was as  low  as 4% of the control and as f.igh  ..
as 57% in another test. The duration of  the exposure did not seem to
have a direct relationship to the survival  totals' but the maximum number
was assumed to be controlled by chamber size.
Van Horn, Willis M., J.  B.  Anderson,  and Max Katy.  1949.  The effect of
     Kraft pulp mill wastes on some aquatic organisms.  Trans. Amer.
     Fish. Soc., 79:55-63.

          Van Horn,  Anderson,  and Katy (1949) determined the minimum
lethal concentration of  hydrogen sulfide for Daphnia sp. to be 1.0 ppra,
for mayfly larvae of the genus Blasturus and Leptophlebia 1.0, and midge
larvae (Chironomus sp.)  750.0  for a period of 48 hours in water with a pli
of 7.6-7.8, and total alkalinity of from 140 to 160 ppm.
                                   30

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          Van Horn, Anderson,  and  Katy  (1949)  determined  the  minimum
lethal concentration of sodium sulfide  for  Daphnia  sp.  to be  10.0  ppm,
for mayfly larvae of the genus Blasturus  and Leptophlebia 1.0 ppm,  ^r.d
midge larvae (Chironomua sp.)  1,000.0 ppn for  a  period  of 48  hours  in
water of pH 7.6-7.8, with total alkalinity  of  from  140  to 160 ppm.
Oseid, Donavon M. and Lloyd L.  Smith,  Jr.   1975.   Long-term effects of
     hydrogen sulfide on Hexagenia  limbata  (Ephemeroptera).   Environ.
     Entomol., 4(1):15-18.

          Oseid and Smith (1975)  determined the 96-hour LC50 of H2S in
acute tests to Hexagenia limbata  nymphs  to  be  0.165 mg/1,  with a water
temperature of 17.8°C to 18.3°C,  dissolved  oxygen 4.5 to 6.6 mg/1,  and pH
of 7.6-7.9.
Smith, Lloyd L.,  Jr. and Donavon M.  Oseid.   1975.   Chronic effects of low
     levels of hydrogen suliide on freshwater fish.   Progress in Water
     Technology,  7(3/4):599-605.

          Smith and Oseid (1975).  Gammarus pseudolimnaeus was subjected
wO-subacute levels of.hydrogen sulfide of 0.0007-0.0153 mg/1, tempera-
tures of 17.1-17.7°C, and pH of 7.7-7.9 for 65,  95,  and 105 days.  At the
higher levels, there was a marked inhibition of  reproduction but at lower
levels stimulation or no differences occurred.   With concentrations of
0.0128 and 0.0153 mg/1 t^S, reproduction  did not maintain the original
population numbers.

          Acute toxicity.tests of hydrogen sulfide to Gammarus pseudolimnaeus
were conducted at a temperature of 18°C,  resulting in a 96-Uour LCso mean
of 0.0220 mg/1 H2S.


Oseid, Uonnvon M. and Lloyd L. Smith, Jr.  1975.  Long-term effects of
     hydrogen sulfide on Hexagenia limbata (Ephemeroptera).  Environ.
     Entomol., 4(1):15-18.

          Oseid and Smith (1975) determined the  12-day LCso i'1 acute
tests to H. limbata nymphs to be 0.060 mg/1, with same water parameters
as above.

          Oseid and Smith (1975) in chronic tests of H. limbata, running
138 days with 0.029 mg/1 H2S, 37% mortality occurred, and at 0'.0762 mg/1,
none survived.  No subimagos emerged at concentrations of 0.0348 mg/1
H2S, but below this level, 30-70% emerged.


Colby, Peter J. and Lloyd L. Smith, Jr.  1967.   Survival of v/alleye eggs
     and fry on paper fiber sludge deposits in Rainy River, Minnesota.
     Trans. Amer. Fish. Soc. 96(3):278-286.
                                    31

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          Colby and Smith (1967)  found that oligorliaet.es and chironomids
were the only groups of bottom organisms  frequently found on wood fiber
deposits with high sulfide concentrations and oxygen deficiencies.

          Colby and Smith (1967)  found 16%, 80%, and 88% mortality of
Gammarus pseudolimnaeus during 24-, 48-,  and 72-hour test.periods in
dissolved sulfide concentrations  of 0.34  ppm.
Oseid, Donavon M. and Lloyd L. Smith, Jr.  1974.  Chronic toxicity of
     hydrogen sulfide to Gammarus pseudolimnaeus.  Trans. Araer. Fish.
     Soc., 4:819-822.

          Oseid and  Smith  (1974) showed  that  in  tests conducted on Gammarus
pseudolimnaeus with  concentrations of H2S below  0.002 mg/1 there was
increased reproduction or  survival in some tests but there was no consistent
effect on mean weight of individuals.  At levels in excess of 0.002 mg/1,
there was a  reduction in numbers and a consequent reduction in total
weight, of the test group.

          Treatments between 0.013 and 0.019  mg/1 showed a mean reduction
of 71% in numbers and 76%.in total weight when compared to the controls.
Oseid, Donavo". M.  and  Lloyd  T,. Smith, Jr.   1975.  Long-term effects of
     hydrogen sulfide  on Hexagenia  limbata  (Eiihemeroptera).  Environ.
     Enromol., 4(1):15-18.

           Oseid  and  Smith  (1975)  showed  in  138-day  chronic tests with the
nymphal  stage of the mayfly  Hex^enis limbata mortality was low--(0-9%) in
all K23  concentrations lower than 0.0290 mg/1.  At  0.0290 cg/1, mortality
was 37%  and  at 0.0762  mg/1 H2S, -none-survived.  Th-ere~was-no~signifi'cant
•reduction  in length  of subimagos  as the  H2S concentration  increased in
each experiment.   Nymphs exposed  to 0.0152  mg/1 H2S were 6% shorter and
at 0.0129  and 0.0290 mg/1  H2S were  3% shorter than  the controls.  The
percentage of subimagos emerging  varied  from 30-75% at levels below
0.0348 mg/1  H2S.   At this  concentration  and higher, no emergence
occurred.
                                     32

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